Part:BBa_K389011:Design

VirA screening device

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

• medium strong constitutive promoter
• The virG gene is mutated so it works without an rpoA subunit from Agrobacterium tumefaciens (YC Jung et al., 2004)
• double terminator (forward) to keep expression of kanamycin resistance by the constitutive promoter low
  • double terminator BBa_B0017 instead of BBa_B0015 because of problems mentioned with BBa_B0012
• Kanamycin resistance for screening with agar plates - the more the vir promoter is activated by VirG the more resistant the cells become to kanamycin.

Source

• virG gene synthesized by Mr. Gene (BBa_K389002)
• vir promoter from A. tumefaciens C58 (BBa_K389003)
• kanR gene made from BioBrick BBa_P1003
• constitutive promoter and double terminator from parts.igem (BBa_J23110, BBa_B0017)

References

YC Jung et al. (2004) Mutants of Agrobacterium tumefaciens virG Gene That Activate Transcription of vir Promoter in Escherichia coli, Current Microbiol 49:334-340.